Cultivation of myocardial tissue by using c-kit+ bone marrow mesenchymal stem cells and decellularized heart matrix / 中国组织工程研究

ABSTRACT

BACKGROUND:Studies have found that c-kit+ bone marrow mesenchymal stem cels can differentiate into myocardial cels specificaly, which may be the ideal seed cels. OBJECTIVE:To investigate the feasibility of cultivation of myocardial tissue by using c-kit+ bone marrow mesenchymal stem cels and decelularized heart matrix. METHODS:Heart tissues harvested from adult rats were decelularized for the folowing experiments. Primary rat bone marrow-derived mesenchymal stem cels were culturedin vitro. Until passage 8, bone marrow mesenchymal stem cels were enriched for c-kit and induced by 5 μmol/L 5-azacytidine for 2 weeks, and a second enrichment for the dihydropyridine receptor subunit α2δ1 was performed before analysis of cardiac differentiation or implantation into decelularized heart matrix for cultivation of myocardial tissue. Six weeks later, myocardial differentiation was identified by specific cardiac protein and action potential. Immunofluorescence staining was used to analysis neonatal myocardial tissue. RESULTS AND CONCLUSION:Six weeks after the second enrichment, 60% bone marrow mesenchymal stem cels expressed cardiac troponin T, GATA binding protein 4, and connexin 43, and these cels could be induced to yield cardiac action potential, which was identified as cardiac differentiation. And when implanted into decelularized heart matrix, these cels could form myocardial tissue arranged regularly.